BMD-PCR技术在新型冠状病毒核酸快速检测中的应用

Application of BMD-PCR technology in rapid detection of 2019-nCoV nucleic acid

目的建立一种可在基层实验室推广的灵敏、快速的新型冠状病毒(2019-nCoV)核酸检测方法。方法利用bubble mediated nucleic acid denaturation(BMD)-PCR技术和2019-nCoV特异性引物及探针建立新型冠状病毒核酸快速检测方法,并利用实验室保存的60份2019-nCoV核酸阳性和68份阴性临床样本对该方法和已批准上市的2019-nCoV核酸检测试剂盒进行比较研究,同时利用实验室保存的11种常见的涵盖常见基因型的经呼吸道和肠道传播的病毒验证该方法的特异性。结果BMD-PCR方法在2019-nCoV核酸检测中具有良好的特异性,与常见的11种经呼吸道和肠道传播的病毒无交叉反应,最低检出限低于600拷贝/ml。与已批准上市的2019-nCoV核酸检测试剂盒相比,128份临床样品的定性检测结果一致性为100%,且检测时间缩短至28 min。结论BMD-PCR 2019-nCoV核酸快速检测可以对临床样品进行快速诊断和鉴定,可以满足目前大规模人群、高通量筛查的紧急需求,具有重要的公共卫生意义。

Objective To establish a sensitive and rapid nucleic acid detection method for 2019-nCoV that can be promoted in primary laboratories.Methods The detection method for 2019-nCoV nucleic acid was established based on the Bubble Mediated Nucleic Acid Denaturation(BMD)-PCR technology with 2019-nCoV specific primers and probes.To validate this method,the laboratory-confirmed clinical samples(including 60 positives and 68 negatives of 2019-nCoV)were used and meanwhile the result were compared with the commercialized 2019-nCoV nucleic acid detection kits.The specificity of this method was validated by using 11 respiratory viruses and enteroviruses which cover common genotypes.Results The BMD-PCR method showed good specificity in the detection of 2019-nCoV nucleic acid.It had no cross-reactivity with 11 common respiratory viruses and enteroviruses,and the limit of detection was less than 600 copies/ml.In addition,the qualitative result of 128 clinical samples were 100%consistent with the commercialized detection kits.Furthermore,the detection time can be shortened to 28 minutes.Conclusions BMD-PCR for 2019-nCoV nucleic acid rapid detection could quickly diagnose and identify clinical samples,and could meet the current urgent needs of large-scale population and high-throughput screening which has important public health significance.